Five-time champion Jimmie Johnson will start from third place, and will be joined on the second row by Championship points leader Greg Biffle.
"Our qualifying effort this year has been improving week in and week out," said Johnson. "I can't express enough how important that is on pit road and late in the race. It just opens up so many options for you on two versus four tyres.
"I'm very proud of the effort today," he added. "I wish we were a little faster but those RPM guys seem to have a little chunk of speed on the field all day long," he said, pointing out that Ambrose had earlier set the top time in practice as well. "I'm happy for those guys, it's a smaller team that is working hard and I know it's a big day for them. I wish it was us, but we will come back and learn a little more Saturday and go racing.”
The drivers get Friday off but have two full practice sessions on Saturday, before the race itself on Sunday evening with the green flag scheduled for shortly after 5.30pm local time (10.30pm BST).
Danica Patrick, having opted to forego any one-off return to open wheel racing at this year's Indianapolis 500 held earlier on the same day, will start from 40th position after making the grid based on car owner points.
That's one row ahead of Kurt Busch, who was the biggest casualty of qualifying after hitting the wall in turn 2 and then rebounding into a hard nose-first hit on the inside wall on his qualifying run, meaning he will need to race the back-up car for the weekend.
"Sorry, guys," radioed Busch, who like Danica had to rely on car owner points to make the starting grid. Trevor Bayne was the highest-placed driver in 11th to qualify on speed alone. Missing out on a Sunday shift were David Reutimann, JJ Yeley, Mike Bliss and Scott Riggs.
Full qualifying and Thursday practice